Monitoring extra-vascular migratory metastasis (EVMM) of migrating cancer cells using an in vitro co-culture system

The mode in which cancer cells migrate away from the primary tumor is not fully understood.In conjunction with intra-vascular cellular migration, we recently proposed extra-vascular migratory metastasis \(EVMM) as a means for cancer cells to venture away from the primary tumor via crawling along the abluminal vascular surface.Here, we propose a protocol that builds on a previous in vitro angiogenesis assay.This protocol details a co-culture approach to monitor uorescence-tagged migrating cancer cells in the presence of vascular structures.This uorescence based single cell coculture approach to monitor cancer cell migratory behavior in real time provides a quantitative approach to decipher the cell decision making processes that cancer cells engage in when undergoing phenotypic switching and exhibiting metastasis like dynamics.This assay can also be adapted for high-throughput analysis and for therapeutic screening of cancer cell metastasis targets